Beijing National Laboratory for Molecular Sciences, Institute of Analytical Chemistry, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China.
Anal Bioanal Chem. 2013 Aug;405(21):6629-38. doi: 10.1007/s00216-013-7109-5. Epub 2013 Jun 18.
An improved online two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry (2D LC-QToF MS) system was developed for the lipid profiling of human plasma, in which different lipid classes were separated by the first dimensional normal-phase (NP) LC and different lipid molecular species were separated by the second dimensional reversed-phase (RP) LC. This 2D LC-QToF MS system was built based on a ten-port, two-position valve as the interface, the conditions of which had been optimized and discussed in detail. As two loops were used to trap and transfer the first dimensional elute to the second dimension separately, this new interface suppressed the sample band broadening in the first dimensional column, increased the recovery and repeatability of 2D LC interface, and offered the possibility for the realization of not-stop-flow NP/RP 2D LC system. Finally, 190 endogenous lipid species out of 10 lipid classes were determined within a single run from the plasma of peritoneal patients. This method was also applied to identify the difference in lipid profile between plasma from peritoneal dialysis patients with bad volume status and peritoneal dialysis patients with good volume status. The discovery of 30 potential biomarkers would be helpful to the malnutrition, inflammation, and atherosclerosis syndrome investigation.
建立了一种改进的在线二维液相色谱-四极杆飞行时间质谱（2D LC-QToF MS）系统，用于人血浆的脂质分析，其中第一维正相（NP）LC 分离不同的脂质类别，第二维反相（RP）LC 分离不同的脂质分子种类。该 2D LC-QToF MS 系统基于十通二位阀作为接口构建，详细优化并讨论了其条件。由于使用两个环分别对第一维洗脱液进行捕获和转移到第二维，该新接口抑制了第一维柱中样品带的展宽，提高了 2D LC 接口的回收率和重现性，为实现不停流 NP/RP 二维 LC 系统提供了可能。最终，从腹膜患者的血浆中单次运行即可确定 10 种脂质类别中的 190 种内源性脂质物质。该方法还应用于鉴定容量状态不佳的腹膜透析患者与容量状态良好的腹膜透析患者之间的血浆脂质谱差异。发现 30 个潜在的生物标志物有助于营养不良、炎症和动脉粥样硬化综合征的研究。
